Western blot detection of the mCP of KSHV from supernatants of KSHV-infected cells in normoxia (21% O2) and hypoxia (1% O2) in the absence or presence of TPA induction.
(A) 96-hour supernatants were collected from BC-3 or BCBL-1 cells that had been cultured for at least 3 passages at 21% O2 or 1% O2, washed, seeded at 300 000 cells per milliliter, and returned to their respective conditions. For BC-3, the band represents approximately 5.5 × 107 viral particles per milliliter of supernatant. For BCBL-1, the band represents approximately 10 × 107 viral particles per milliliter. These values are based on comparison with a viral standard electrophoresed on the same blot. (B) BCBL-1 cells cultured at 300 000 cells per milliliter in normoxia (21% O2) or acute hypoxia (1% O2) were then treated with 6.4 nM TPA. The time indicated represents the time following TPA addition after incubation of cells overnight. Each band was scanned, and the relative band intensity expressed as total pixels was determined and plotted for samples in normoxia (▨ and top band) and hypoxia (▪ and bottom band).
